Rapid determination of total acetylation levels in cells ------- Immunoaffinity chromatography-ELISA method
Step 1: Purification of acetylated protein by affinity chromatography
1. Cleavage the cells with cell lysate, centrifuge at 10,000 rpm for 2 minutes, and take the supernatant for later use.
2. Take 20-40 ul of anti-acetylated lysine Agarose (ICP0388, as shown on the right).
Wash twice with PBS, 1 ml each time.
3. Mix the cell lysis supernatant with the washed packing (ICP0388) and incubate for 60 minutes at a low speed on a 39oC rotary shaker.
4. Centrifuge at 1000 rpm for 1 minute, remove the supernatant, resuspend the pellet with PBSt, centrifuge at 1000 rpm for 1 minute, remove the supernatant, and repeat 3 times.
Wash with PBS for the last time and discard the supernatant.
Note: Cell lysates should contain protease inhibitors, histone acetylase inhibitors, etc.
Step 2: release and coating to separate and purify the purified acetylated protein
5. Boil the anti-lysine antibody filler-acetylated protein complex of step 2 in boiling water for 2 minutes with 20ul 0.1M HCL, 10000rpm
After centrifugation for 1 minute, the supernatant liquid was completely transferred to an ELISA plate containing 80 ul of 0.2 M Na2CO3, and incubated at 39 ° C for 30 minutes;
6. Remove the ELISA plate and pat dry. Wash twice with PBSt and pat dry each time. Add 100ul of PBSt with 0.5% BSA, 39oC
Closed for 10 minutes.
Step 3: Detection of protein acetylation levels with anti-acetyl lysine antibody HRP markers
7. Remove the ELISA plate, pat dry, wash twice with PBSt, and pat dry. Add 100ul of 0.25ug/ml anti-lysine antibody
The HRP-conjugated complex (Anti-acetylated lysine HRP conugates, ICP0381, as shown on the right) was incubated at 39oC for 30 minutes.
8. Remove the ELISA plate, pat dry, and clean the plate well 3 times with PBSt, each time patted dry. Add 100ul of TMB solution and incubate at 39oC
10-20 minutes. The reaction was stopped by the final addition of 100 ul of 0.1 M H2SO4 and read at a wavelength of 450 nm.
The Infrared Thermometer is designed for measuring the human body forehead temperature reference. It is very simple and convenient to use. Accurate temperature measurement in 1 second, no laser spot, avoid potential damage to eyes, no need to touch human skin, avoid cross infection, one-click temperature measurement, and check for flu. It is suitable for home users, hotels, libraries, large enterprises and institutions, and can also be used in hospitals, schools, customs, airports and other comprehensive places, and can also be provided to medical personnel for use in clinics.
The normal body temperature of the human body averages between 36 ~ 37 ℃), exceeding 37.1 ℃ means fever, 37.3_38 ℃ is low fever, and 38.1_40 ℃ is high fever. Life above 40 ° C is always dangerous.
1. Human body temperature measurement: accurate measurement of human body temperature, replacing traditional mercury thermometers. Women who want to have a child can use an infrared thermometer (frontal temperature gun) to monitor the basal body temperature at any time, record the body temperature during ovulation, and choose the right time to conceive, and can also measure the temperature to judge pregnancy.
Of course, the most important thing is to observe whether your body temperature is abnormal at any time, to avoid flu infection, and to prevent swine flu.
2. Skin temperature measurement: To measure the surface temperature of the human skin, for example, it can be used for amputation and re-implantation surgery.
3. Object temperature measurement: To measure the surface temperature of an object, for example, it can be used to measure the temperature of the surface of a tea cup.
4. Liquid temperature measurement: measure the temperature of the liquid, such as the temperature of the baby's bath water. When the baby takes a bath, measure the water temperature, and no longer worry about being cold or hot; you can also measure the water temperature of the milk bottle to facilitate the preparation of Baby's milk powder
5. Room temperature can be measured:
※Precautions:
1. Please read the instructions carefully before the measurement, and the forehead should be kept dry, and the hair should not cover the forehead (please carry out the measurement in the environment of 10 ℃ -40 ℃) to ensure the accuracy of the measurement.
2. The forehead temperature quickly measured by this product is for reference only, and should not be used as a basis for medical judgment. If you find abnormal body temperature, please use a medical thermometer for further measurement.
3. Please protect the sensing lens and clean it in time. If the change of the ambient temperature is too large, you need to place the measuring instrument in the environment to be measured for 20 minutes, wait for it to adapt to the ambient temperature, and then use it to measure more accurate values.
Infrared Thermometer
Reusable Infrared Thermometer,Noncontact Infrared Thermometer,Reusable Medical Infrared Thermometer,Medical Noncontact Infrared Thermometer
Guangzhou Aikangli Medical Technology Co., Ltd. , https://www.aikanli.com